WO2024093907A1 - Ligand pour administrer de l'arnsi à l'oeil et au système nerveux central - Google Patents

Ligand pour administrer de l'arnsi à l'oeil et au système nerveux central Download PDF

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WO2024093907A1
WO2024093907A1 PCT/CN2023/127728 CN2023127728W WO2024093907A1 WO 2024093907 A1 WO2024093907 A1 WO 2024093907A1 CN 2023127728 W CN2023127728 W CN 2023127728W WO 2024093907 A1 WO2024093907 A1 WO 2024093907A1
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compound
nhc
alkyl
formula
mmol
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Chinese (zh)
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黄金宇
方剑武
杨晓彦
邹昊
刘俊凯
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大睿生物医药科技(上海)有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
    • C07D265/28—1,4-Oxazines; Hydrogenated 1,4-oxazines
    • C07D265/30—1,4-Oxazines; Hydrogenated 1,4-oxazines not condensed with other rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Definitions

  • the present invention belongs to the field of medicine, and specifically relates to a hydrophobic group having the ability to enhance the ability of double-stranded RNA to pass through the blood-brain barrier and/or be delivered to the eye, such as the R group in formula (I), and a compound of formula (I) in which the hydrophobic group is connected to a nucleotide, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • RNA interference is a phenomenon in which double-stranded RNA (dsRNA, also known as siRNA) induces efficient and specific degradation of target mRNA.
  • dsRNA double-stranded RNA
  • siRNA siRNA
  • siRNA due to the presence of the blood-brain barrier, it is difficult to deliver siRNA to the central nervous system to exert its effect, which limits the application of siRNA.
  • Some attempts have been made in the art to deliver siRNA to the central nervous system such as WO2004094595A2, which discloses the use of a single lipid ligand (such as cholesterol or long-chain alkane) at the end of the chain to deliver siRNA, WO2019217459A1, which discloses the use of a single lipid ligand to deliver siRNA inside the chain, and WO2021092371A2, which discloses a series of new lipid ligand structures.
  • the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • the present invention provides an oligonucleotide comprising one or more compounds of formula (I'), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • the present invention provides a double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, the antisense strand comprising a sequence that is sufficiently complementary to the sense strand and the target mRNA, wherein the sense strand and/or the antisense strand comprises one or more compounds of formula (I'), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • hydrophobic group (R group in formula (I)) provided by the present invention can be linked to the siRNA via an existing linker structure, such as a biodegradable linker structure.
  • the present invention provides a vector comprising a nucleotide sequence encoding the aforementioned double-stranded RNA.
  • the present invention provides a cell containing the aforementioned double-stranded RNA or the aforementioned vector.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the aforementioned double-stranded RNA, the aforementioned vector, or the aforementioned cell, and optionally a pharmaceutically acceptable carrier or excipient.
  • the present invention provides a kit comprising the aforementioned double-stranded RNA, the aforementioned vector, or the aforementioned cell.
  • FIG1 shows the reduction in the expression of SOD1 in the cervical spinal cord, thoracic spinal cord, cerebellum, brainstem, hippocampus and frontal cortex after puncture and injection of the siRNA of the present invention in SD rats.
  • FIG. 2 shows the reduction in the expression of the TTR gene in the eyes of C57BL/6 mice after bilateral intravitreal injection of the siRNA of the present invention.
  • C 1-6 alkyl includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 1-6 , C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-5 , C 2-4 , C 2-3 , C 3-6 , C 3-5 , C 3-4 , C 4-6 , C 4-5 , and C 5-6 alkyl.
  • C 1-30 alkyl refers to a straight or branched saturated hydrocarbon group having 1 to 30 carbon atoms. In some embodiments, C 5-25 alkyl, C 10-20 alkyl, C 1-20 alkyl, C 1-10 alkyl and C 1-6 alkyl are preferred.
  • C 1-6 alkyl examples include: methyl (C 1 ), ethyl (C 2 ), n-propyl (C 3 ), isopropyl (C 3 ), n-butyl (C 4 ), tert-butyl (C 4 ), sec-butyl (C 4 ), isobutyl (C 4 ), n-pentyl (C 5 ), 3-pentyl (C 5 ), pentyl (C 5 ), neopentyl (C 5 ), 3-methyl-2-butyl (C 5 ), tert-pentyl (C 5 ) and n-hexyl (C 6 ).
  • C 1-6 alkyl also includes heteroalkyl groups in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus).
  • the alkyl group may be optionally substituted by one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.
  • alkyl abbreviations include: Me(-CH 3 ), Et(-CH 2 CH 3 ), iPr(-CH(CH 3 ) 2 ), nPr(-CH 2 CH 2 CH 3 ), n-Bu(-CH 2 CH 2 CH 2 CH 3 ) or i-Bu(-CH 2 CH(CH 3 ) 2 ).
  • C2-30 alkenyl refers to a straight or branched hydrocarbon group having 2 to 30 carbon atoms and at least one carbon-carbon double bond. In some embodiments, C10-25 alkenyl, C2-10 alkenyl, C2-6 alkenyl and C2-4 alkenyl are preferred. Examples of C2-6 alkenyl include: vinyl ( C2 ), 1-propenyl ( C3 ), 2-propenyl ( C3 ), 1-butenyl ( C4 ), 2-butenyl (C4), butadienyl ( C4 ), pentenyl ( C5 ), pentadienyl ( C5 ), hexenyl ( C6 ), and the like.
  • C2-6 alkenyl also includes heteroalkenyl, wherein one or more (e.g., 1 , 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus).
  • An alkenyl group can be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.
  • C 2-30 alkynyl refers to a straight or branched hydrocarbon group having 2 to 30 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. In some embodiments, C 10-25 alkynyl, C 2-10 alkynyl, C 2-6 alkynyl and C 2-4 alkynyl are preferred.
  • C 2-6 alkynyl examples include, but are not limited to, ethynyl (C 2 ), 1-propynyl (C 3 ), 2-propynyl (C 3 ), 1-butynyl (C 4 ), 2-butynyl (C 4 ), pentynyl (C 5 ), hexynyl (C 6 ), and the like.
  • C 2-6 alkynyl also includes heteroalkynyl groups in which one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus).
  • An alkynyl group can be optionally substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.
  • C 1-10 alkylene refers to a divalent group formed by removing another hydrogen of a C 1-10 alkyl, a C 2-10 alkenyl and a C 2-10 alkynyl, respectively, and may be substituted or unsubstituted.
  • C 2-8 alkylene, C 3-7 alkylene, C 4-6 alkylene, C 1-4 alkylene, C 2-4 alkylene and C 1-3 alkylene are preferred.
  • Unsubstituted alkylene includes, but is not limited to, methylene (—CH 2 —), ethylene (—CH 2 CH 2 —), propylene (—CH 2 CH 2 CH 2 —), butylene (—CH 2 CH 2 CH 2 CH 2 —), pentylene (— CH 2 CH 2 CH 2 CH 2 CH 2 —), hexylene (—CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 —), and the like.
  • substituted alkylene groups for example, substituted alkylene groups with one or more alkyl(methyl) groups, include, but are not limited to, substituted methylene groups (—CH(CH 3 )—, —C(CH 3 ) 2 —), substituted ethylene groups (—CH(CH 3 )CH 2 —, —CH 2 CH(CH 3 )—, —C(CH 3 ) 2 CH 2 —, —CH 2 C(CH 3 ) 2 — ), substituted propylene groups (—CH(CH 3 )CH 2 CH 2 —, —CH 2 CH(CH 3 )CH 2 —, —CH 2 CH 2 CH(CH 3 )—, —C(CH 3 ) 2 CH 2 CH 2 —, —CH 2 C(CH 3 ) 2 CH 2 —, —CH 2 CH 2 C(CH 3 ) 2 —), and the like.
  • substituted methylene groups —CH(CH 3 )—, —C(CH 3 ) 2 —
  • C 0-10 alkylene group means a chemical bond as well as the above-mentioned "C 1-10 alkylene group”.
  • Halo or "halogen” refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).
  • C 1-20 haloalkyl refers to the above-mentioned “C 1-20 alkyl”, “C 1-6 alkyl” and “C 1-4 alkyl", respectively, which are substituted with one or more halogen groups.
  • C 1-4 haloalkyl is particularly preferred, and C 1-2 haloalkyl is more preferred.
  • haloalkyl groups include, but are not limited to, -CF 3 , -CH 2 F, -CHF 2 , -CHFCH 2 F, -CH 2 CHF 2 , -CF 2 CF 3 , -CCl 3 , -CH 2 Cl, -CHCl 2 , 2,2,2-trifluoro-1,1-dimethyl-ethyl, and the like.
  • the haloalkyl group can be substituted at any available attachment point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.
  • hydrophobic group refers broadly to any chemical group that has an affinity for lipids.
  • One way to characterize the hydrophobicity of a hydrophobic group is by the octanol-water partition coefficient logKow , where Kow is the ratio of the concentration of a chemical in the octanol phase to its concentration in the aqueous phase when a two-phase system is at equilibrium.
  • logKow of the hydrophobic moiety is greater than 1, greater than 1.5, greater than 2, greater than 3, greater than 4, greater than 5, or greater than 10.
  • the hydrophobic moiety is the R group in the compound of formula I.
  • Alkyl, alkenyl, alkynyl, etc. as defined herein are optionally substituted groups.
  • each of Raa is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, or two Ra groups combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;
  • each of R bb is independently selected from the group consisting of hydrogen, -OH, -OR aa , -N(R cc ) 2 , -CN, -C( ⁇ O)R aa , -C( ⁇ O)N(R cc ) 2 , -CO 2 R aa , -SO 2 R aa , -C( ⁇ NR cc )OR aa , -C( ⁇ NR cc )N(R cc ) 2 , -SO 2 N(R cc ) 2 , -SO 2 R cc , -SO 2 OR cc , -SOR aa , -C( ⁇ S)N(R cc ) 2 , -C( ⁇ O)SR cc , -C( ⁇ S)SR cc , -P( ⁇ O) 2 R aa , -P( ⁇ O)(R aa ) 2
  • each of R cc is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R cc groups combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 R dd groups;
  • Each of R ee is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, and heteroaryl, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R gg groups;
  • each of Rff is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two Rff groups combine to form a heterocyclyl or heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently substituted with 0, 1, 2, 3, 4 or 5 Rgg groups;
  • siRNA refers to a class of double-stranded RNA molecules that can mediate the silencing of a target RNA (e.g., mRNA, e.g., a transcript of a gene encoding a protein) that is complementary thereto.
  • a target RNA e.g., mRNA, e.g., a transcript of a gene encoding a protein
  • siRNA is typically double-stranded, comprising an antisense strand complementary to the target RNA, and a sense strand complementary to the antisense strand.
  • mRNA is also referred to herein as mRNA to be silenced.
  • a gene is also referred to as a target gene.
  • the RNA to be silenced is an endogenous gene or a pathogen gene.
  • RNA (e.g., tRNA) and viral RNA other than mRNA can also be targeted.
  • antisense strand refers to a strand of an siRNA that includes a region that is completely, fully or substantially complementary to a target sequence.
  • sense strand refers to a strand of an siRNA that includes a region that is completely, fully or substantially complementary to a region that is the antisense strand as defined herein.
  • complementary region refers to a region on the antisense strand that is completely, fully or substantially complementary to the target mRNA sequence.
  • mispairing can be located in the interior or terminal regions of the molecule.
  • the most tolerated mispairing is located in the terminal regions, for example, within 5, 4, 3, 2 or 1 nucleotides at 5' and/or 3' ends.
  • the antisense strand portion that is most sensitive to mispairing is referred to as a "seed region".
  • seed region For example, in a siRNA comprising a 19nt chain, the 19th position (from 5' to 3') can tolerate some mispairing.
  • complementary refers to the ability of a first polynucleotide to hybridize to a second polynucleotide under certain conditions, such as stringent conditions.
  • stringent conditions may include 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mM EDTA at 50°C or 70°C for 12-16 hours.
  • “complementary” sequences may also include or be formed entirely from non-Watson-Crick base pairs and/or base pairs formed from non-natural and modified nucleotides.
  • Such non-Watson-Crick base pairs include, but are not limited to, G:U wobble base pairing or Hoogstein base pairing.
  • a polynucleotide that is "at least partially complementary,” “fully complementary,” or “substantially complementary” to a messenger RNA (mRNA) refers to a polynucleotide that is substantially complementary to a contiguous portion of an mRNA of interest.
  • mRNA messenger RNA
  • a polynucleotide is complementary to at least a portion of a PCSK9 mRNA if the sequence is substantially complementary to a non-interrupted portion of an mRNA encoding PCSK9.
  • Fully complementary refers to the extent to which the sense strand only needs to be complementary to the antisense strand in order to maintain the overall double-stranded characteristics of the molecule.
  • perfect complementarity is generally required, in some cases, particularly in the antisense strand, one or more, such as 6, 5, 4, 3, 2 or 1 mismatches (relative to the target mRNA) may be included, but the sense strand and the antisense strand can still maintain the overall double-stranded characteristics of the molecule.
  • shRNA refers to short hairpin RNA.
  • shRNA includes two short inverted repeat sequences.
  • the shRNA cloned into the shRNA expression vector includes two short inverted repeat sequences separated by a stem-loop sequence in the middle, forming a hairpin structure controlled by the polIII promoter. Then 5-6 Ts are connected as the transcription terminator of RNA polymerase III.
  • Nucleoside is a compound composed of two substances, a purine base or a pyrimidine base, and ribose or deoxyribose
  • nucleotide is a compound composed of three substances, a purine base or a pyrimidine base, ribose or deoxyribose, and phosphate
  • oligonucleotide refers to a nucleic acid molecule (RNA or DNA) with a length of, for example, less than 100, 200, 300 or 400 nucleotides.
  • Base is the basic unit of synthesis of nucleosides, nucleotides and nucleic acids. It contains nitrogen and is also called “nitrogenous base”.
  • capital letters A, U, T, G and C represent the base composition of nucleotides, which are adenine, uracil, thymine, guanine and cytosine respectively.
  • the "modification" of the nucleotides described herein includes, but is not limited to, methoxy modification, fluorination modification, phosphorothioate linkage, or conventional protecting group protection, etc.
  • the fluorination-modified nucleotide refers to a nucleotide in which the 2'-hydroxyl group of the ribose group of the nucleotide is replaced by fluorine
  • the methoxy-modified nucleotide refers to a nucleotide in which the 2'-hydroxyl group of the ribose group is replaced by a methoxy group.
  • Modified nucleotides herein include, but are not limited to, 2'-O-methyl modified nucleotides, 2'-fluoro modified nucleotides, 2'-deoxy-modified nucleotides, inosine ribonucleotides, abasic nucleotides, reverse abasic deoxyribonucleotides, nucleotides containing thiophosphate groups, vinyl phosphate modified nucleotides, locked nucleotides, 2'-amino-modified nucleotides, 2'-alkyl-modified nucleotides, morpholino nucleotides, phosphoramidates, non-natural bases containing nucleotides, and terminal nucleotides, deoxyribonucleotides or conventional protective groups connected to cholesterol derivatives or dodecanoic acid didecylamide groups.
  • the 2'-fluoro modified nucleotide refers to a nucleotide in which the hydroxyl group at the 2' position of the ribose group of the nucleotide is replaced by fluorine.
  • the 2'-deoxy-modified nucleotide refers to a nucleotide in which the 2'-hydroxyl group of the ribose group is replaced by a methoxy group.
  • Ligand moiety refers to a chemical moiety conjugated to a siRNA that is capable of altering the distribution, targeting or lifetime of the siRNA.
  • a ligand provides enhanced affinity for a selected target (e.g., a molecule, a cell or cell type, a compartment (e.g., a cell or organ compartment, a tissue, an organ, or a region of the body) compared to, for example, a siRNA in the absence of such a ligand.
  • Reactive phosphorus group refers to a phosphorus-containing group contained in a nucleotide unit or a nucleotide analog unit, which can react with a hydroxyl or amine group contained in another molecule, especially in another nucleotide unit or in another nucleotide analog, by a nucleophilic attack reaction. Typically, such a reaction produces an ester-type internucleoside bond connecting the first nucleotide unit or the first nucleotide analog unit to the second nucleotide unit or the second nucleotide analog unit.
  • the reactive phosphorus group can be selected from phosphoramidites, H-phosphonates, alkyl-phosphonates, phosphates or phosphate mimetics, including but not limited to: natural phosphates, thiophosphates, dithiophosphates, borane phosphates, borane thiophosphates, phosphonates, halogen-substituted phosphonates and phosphates, phosphoramidates, phosphodiesters, phosphotriesters, thiophosphorodiesters, thiophosphorothioates, diphosphates and triphosphates, preferably -P(OCH 2 CH 2 CN)(N(iPr) 2 ).
  • Protecting group refers to any atom or group of atoms added to a molecule to prevent an existing group in the molecule from undergoing an undesirable chemical reaction.
  • a “protecting group” may be an unstable chemical moiety known in the art that is used to protect reactive groups, such as hydroxyl, amino, and thiol groups, to prevent undesirable or inappropriate reactions during chemical synthesis.
  • Protecting groups are typically used selectively and/or orthogonally to protect sites during reactions at other reactive sites and can then be removed to leave the unprotected group intact or available for further reactions.
  • a non-limiting list of protecting groups includes benzyl; substituted benzyl; alkylcarbonyl and alkoxycarbonyl (e.g., tert-butyloxycarbonyl (BOC), acetyl or isobutyryl); arylalkylcarbonyl and arylalkoxycarbonyl (e.g., benzyloxycarbonyl); substituted methyl ethers (e.g., methoxymethyl ether); substituted ethyl ethers; substituted benzyl ethers; tetrahydropyranyl ethers; silyl (e.g., trimethylsilyl, triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, tri-isopropylsilyloxymethyl, [2-(trimethylsilyl)ethoxy] methyl or tert-butyldiphenylsilyl); esters (e.g.
  • Preferred protecting groups are selected from acetyl (Ac), benzoyl (Bzl), benzyl (Bn), isobutyryl (iBu), phenylacetyl, benzyloxymethyl acetal (BOM), ⁇ -methoxyethoxymethyl ether (MEM), methoxymethyl ether (MOM), p-methoxybenzyl ether (PMB), methylthiomethyl ether, neopentyl (V), benzyloxymethyl acetal (BOM), benzyloxymethyl ether (BOM ...
  • Hydro protecting group refers to a group that can protect the hydroxyl group from chemical reactions and can be removed under specific conditions to restore the hydroxyl group. It mainly includes silane type protecting groups, acyl type protecting groups or ether type protecting groups, preferably the following:
  • TMS trimethylsilyl
  • TES triethylsilyl
  • DMIPS dimethylisopropylsilyl
  • DEIPS diethylisopropylsilyl
  • TDMS tert-butyldimethylsilyl
  • TDPS tert-butyldiphenylsilyl
  • TIPS triisopropylsilyl
  • acetyl (Ac) chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), benzoyl, p-methoxybenzoyl, 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxycarbonyl (Boc), benzyl (Bn), p-meth
  • pharmaceutically acceptable salt refers to those carboxylates, amino acid addition salts of the compounds of the present invention which are suitable for use in contact with patient tissues within the scope of sound medical judgment, do not produce undue toxicity, irritation, allergic response, etc., are commensurate with a reasonable benefit/risk ratio, and are effective for their intended use, including (where possible) zwitterionic forms of the compounds of the present invention.
  • the present invention includes tautomers, which are functional isomers produced by rapid movement of an atom in two positions in a molecule.
  • tautomers which are functional isomers produced by rapid movement of an atom in two positions in a molecule.
  • Compounds that exist in different tautomeric forms are not limited to any specific tautomer, but are intended to cover all tautomeric forms.
  • the compounds of the present invention may include one or more asymmetric centers and may therefore exist in a variety of stereoisomeric forms, for example, enantiomers and/or diastereoisomeric forms.
  • the compounds of the present invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers.
  • Isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.
  • HPLC high pressure liquid chromatography
  • the present invention also includes isotopically labeled compounds (isotopic variants), which are equivalent to those described in formula (I), but one or more atoms are replaced by atoms having atomic masses or mass numbers different from the atomic masses or mass numbers commonly found in nature.
  • isotopes that can be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F and 36 Cl, respectively.
  • isotopically labeled compounds of formula (I) of the present invention and their prodrugs can generally be prepared by replacing non-isotopically labeled reagents with readily available isotopically labeled reagents when carrying out the processes disclosed in the following schemes and/or the Examples and Preparations.
  • the present invention specifically relates to a compound of formula (I), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • L1 and L2 are independently selected from H, a reactive phosphorus group, a hydroxyl protecting group or a solid support;
  • Rs is selected from H, D, halogen, C1-6 alkyl or C1-6 haloalkyl, which is optionally deuterated until fully deuterated;
  • n 0, 1, 2, 3, 4, 5 or 6;
  • R is -C(O)-C 0-10 alkylene-LR 1 , -C(O)-C 2-10 alkenylene-LR 1 or -C(O)-C 2-10 alkynylene-LR 1 ;
  • L is a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SS-, -NHC(O)O-, -NHC(O)NH-, -OC(O)O-, -OC(O)NH-, -NHC(O)-CH(OR 1 )CH 2 O-, -C(O)NH-CH(OR 1 )CH 2 O-, -OC(O)-CH(OR 1 )CH 2 O-, -C(O)O-CH(OR 1 )CH 2 O-, -NHC(O)-CH(R 1 )-, -C(O)NH-CH(R 1 )-, -OC(O)-CH(R 1 )- , -C(O)O-CH(R 1 )- , -CH(OR 1 )CH 2 O- , -O-CH 2 CH(R 1 )O-,
  • R 1 is independently C 1-30 alkyl, C 2-30 alkenyl or C 2-30 alkynyl, wherein 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 non-adjacent carbon atoms in the group may be replaced by heteroatoms selected from O, S and N, or the -CH 2 CH 2 - group may be replaced by -OC(O)-, -C(O)O-, -NHC(O)- or -C(O)NH-, or the substituents on one or more carbon atoms may be linked to form a saturated or unsaturated ring;
  • hydrogen atoms in the C0-10 alkylene, C2-10 alkenylene, C2-10 alkynylene, C1-30 alkyl, C2-30 alkenyl and C2-30 alkynyl groups may be optionally replaced by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more halogens, C1-6 alkyl or C1-6 haloalkyl groups, and are optionally deuterated until fully deuterated.
  • the present invention also relates to an oligonucleotide comprising one or more compounds of formula (I'), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • L 2 represents H or a solid support, or represents the position of connection with the adjacent nucleotide
  • the present invention also relates to a double-stranded RNA having a sense strand and an antisense strand, each strand having 14 to 30 nucleotides, wherein the antisense strand comprises a sequence that is sufficiently complementary to the sense strand and the target mRNA, wherein the sense strand and/or the antisense strand comprises one or more compounds of formula (I'), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • L 2 represents H or a hydroxyl protecting group, or represents the position for connection with the adjacent nucleotide
  • L1 is H; in another embodiment, L1 is a reactive phosphorus group; in another embodiment, L1 is a hydroxyl protecting group; in another embodiment, L1 is a solid support.
  • L2 is H; in another embodiment, L2 is a reactive phosphorus group; in another embodiment, L2 is a hydroxyl protecting group; in another embodiment, L2 is a solid support; in another embodiment, L2 represents the position of attachment to the adjacent nucleotide.
  • one of L 1 and L 2 is -C(O)CH 2 CH 2 C(O)OH or 4,4'-dimethoxytrityl; in another more specific embodiment, one of L 1 and L 2 is -C(O)CH 2 CH 2 C(O)OH.
  • Rs is H; in another embodiment, Rs is D; in another embodiment, Rs is halogen; in another embodiment, Rs is C 1-6 alkyl; in another embodiment, Rs is C 1-6 haloalkyl; in another embodiment, when Rs is C 1-6 alkyl or C 1-6 haloalkyl, the group is optionally deuterated until fully deuterated;
  • R is -C(O)-C 0-10 alkylene-LR 1 ; in another embodiment, R is -C(O)-C 2-10 alkenylene-LR 1 ; in another embodiment, R is -C(O)-C 2-10 alkynylene-LR 1 .
  • R is -C(O)-LR 1 ; in another more specific embodiment, R is -C(O)-C 2-8 alkylene-LR 1 ; in another more specific embodiment, R is -C(O)-C 3-7 alkylene-LR 1 ; in another more specific embodiment, R is -C(O)-C 4-6 alkylene-LR 1 ; in another more specific embodiment, R is -C(O)-C 1-3 alkylene-LR 1 .
  • L is a chemical bond; in another embodiment, L is -NHC(O)-; in another embodiment, L is -C(O)NH-; in another embodiment, L is -OC(O)-; in another embodiment, L is -C(O)O-; in another embodiment, L is -SS-; in another embodiment, L is -NHC(O)O-; in another embodiment, L is -NHC(O)NH-; in another embodiment, L is -OC(O)O-; in another embodiment, L is -OC(O)NH-; in another embodiment, L is -NHC(O)-CH(OR 1 )CH 2 O-; in another embodiment, L is -C(O)NH-CH(OR 1 )CH 2 O-; in another embodiment, L is -OC(O)-CH(OR 1 )CH 2 O-; in another embodiment, L is -C(O)CH(OR 1 )CH 2 O-; in another embodiment, L is -OC(O)-CH(OR 1
  • L is a bond, -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SS-, -NHC(O)O-, -NHC(O)NH-, -OC(O) O- , -OC(O)NH-, -O-CH(CH(OH)CH2OH)-, -O - CH(CH(NH2)CH2OH)-, -O-CH(CH2OH ) CH ( OH)-, -NH-CH ( CH2OH)CH(OH) - , -O-CH2CH(OH)CH( OH ) - , -NHC(O)-CH2- O -CH(CH(OH)CH2OH)-, -NHC(O) -CH2 -O-CH(CH( NH2 ) CH2OH )-, -NHC(O)-CH2- O -CH(CH(OH)CH2OH)-
  • L is -NHC(O)-CH(OR 1 )CH 2 O-, -C(O)NH-CH(OR 1 )CH 2 O-, -OC(O)-CH(OR 1 )CH 2 O-, -C(O)O-CH(OR 1 )CH 2 O-, -NHC(O)-CH(R 1 )-, -C(O)NH-CH(R 1 )-, -OC(O)-CH(R 1 )-, -C(O)O-CH(R 1 )-, -CH(OR 1 )CH 2 O-, -O-CH(R 1 )CH 2 O-, or -O-CH 2 CH(R 1 )O-; in another more specific embodiment, L is -NHC(O)-CH(OR 1 )CH 2 O-, -NHC(O)-CH(R 1 )-, or -CH(OR 1 )CH 2 O-; in another more specific embodiment, L
  • R 1 is C 1-30 alkyl; in another embodiment, R 1 is C 2-30 alkenyl; in another embodiment, R 1 is C 2-30 alkynyl; in another embodiment, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 non-adjacent carbon atoms in the group in R 1 may be replaced by heteroatoms selected from O, S and N; in another embodiment, the -CH 2 CH 2 - group in R 1 may be replaced by -OC(O)-, -C(O)O-, -NHC(O)- or -C(O)NH-; in another embodiment, the substituents on one or more carbon atoms in R 1 may be connected to form a saturated or unsaturated ring.
  • R 1 is independently C 1-30 alkyl or C 2-30 alkenyl, wherein 1, 2, 3, 4, 5, 6, 7 or 8 non-adjacent carbon atoms in the group may be replaced by heteroatoms selected from O, S and N, or a -CH 2 CH 2 - group may be replaced by -NHC(O)- or -C(O)NH-, or substituents on one or more carbon atoms may be linked to form a saturated or unsaturated ring; in another more specific embodiment, R 1 is independently C 5-25 alkyl, a C 10-25 alkenyl containing 1, 2, 3, 4, 5 or 6 double bonds, a C 5-25 alkyl in which 1, 2, 3, 4 or 5 carbon atoms are replaced by N heteroatoms and/or 1, 2 or 3 -CH 2 CH 2 - groups are replaced by -C(O)NH-, or a C 5-25 alkyl in which substituents on one or more carbon atoms are linked to form a steroidal ring; in another
  • any technical solution or any combination thereof in any of the above specific embodiments can be combined with any technical solution or any combination thereof in other specific embodiments.
  • any technical solution or any combination thereof of L1 can be combined with any technical solution or any combination thereof of L2 , Rs , m, R, L and R1 , etc.
  • the present invention is intended to include all combinations of these technical solutions, which are not listed one by one due to space limitations.
  • the present invention also provides a vector, which comprises a nucleotide sequence encoding the siRNA of the present invention.
  • the vector of the present invention can amplify or express the nucleotide sequence encoding the siRNA of the present invention connected thereto.
  • siRNA targeting the PCSK9 gene can be expressed from a transcription unit inserted into a DNA or RNA vector. Expression can be short-lived (within hours to weeks) or continuous (weeks to months or longer), depending on the specific construct used and the target tissue or cell type.
  • the coding nucleotides of the siRNA can be introduced into a linear construct, a circular plasmid or a viral vector.
  • the nucleotides of the siRNA can be integrated into the cell genome for stable expression, or expressed in a stable extrachromosomal inheritance.
  • siRNA expression vectors are typically DNA plasmids or viral vectors.
  • Viral vector systems containing siRNA coding sequences include but are not limited to: (a) adenovirus vectors; (b) retrovirus vectors; (c) adeno-associated virus vectors; (d) herpes simplex virus vectors; (e) SV40 vectors; (f) polyoma virus vectors; (g) papilloma virus vectors; (h) picornavirus vectors; (i) poxvirus vectors; and (j) helper virus-dependent adenovirus or gut-free adenovirus.
  • the present invention also provides a cell containing the siRNA or vector of the present invention, wherein the siRNA or vector of the present invention can be transcribed in the cell.
  • L1 and L2 are independently selected from H, a reactive phosphorus group, a hydroxyl protecting group or a solid support;
  • Rs is selected from H, D, halogen, C1-6 alkyl or C1-6 haloalkyl, which is optionally deuterated until fully deuterated;
  • n 0, 1, 2, 3, 4, 5 or 6;
  • R is -C(O)-C 0-10 alkylene-LR 1 , -C(O)-C 2-10 alkenylene-LR 1 or -C(O)-C 2-10 alkynylene-LR 1 ;
  • L is a chemical bond, -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SS-, -NHC(O)O-, -NHC(O)NH-, -OC(O)O-, -OC(O)NH-, -NHC(O)-CH(OR 1 )CH 2 O-, -C(O)NH-CH(OR 1 )CH 2 O-, -OC(O)-CH(OR 1 )CH 2 O-, -C(O)O-CH(OR 1 )CH 2 O-, -NHC(O)-CH(R 1 )-, -C(O)NH-CH(R 1 )-, -OC(O)-CH(R 1 )- , -C(O)O-CH(R 1 )- , -CH(OR 1 )CH 2 O- , -O-CH 2 CH(R 1 )O-,
  • R 1 is independently C 1-30 alkyl, C 2-30 alkenyl or C 2-30 alkynyl, wherein 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 non-adjacent carbon atoms in the group may be replaced by heteroatoms selected from O, S and N, or the -CH 2 CH 2 - group may be replaced by -OC(O)-, -C(O)O-, -NHC(O)- or -C(O)NH-, or the substituents on one or more carbon atoms may be linked to form a saturated or unsaturated ring;
  • hydrogen atoms in the C0-10 alkylene, C2-10 alkenylene, C2-10 alkynylene, C1-30 alkyl, C2-30 alkenyl and C2-30 alkynyl groups may be optionally replaced by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more halogens, C1-6 alkyl or C1-6 haloalkyl groups, and are optionally deuterated until fully deuterated.
  • Technical solution 10 The compound of any one of technical solutions 1-9, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein the compound is selected from the following:
  • An oligonucleotide comprising one or more compounds of formula (I'), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof:
  • L 2 represents H or a solid support, or represents the position of connection with the adjacent nucleotide
  • Rs is selected from H, D, halogen, C1-6 alkyl or C1-6 haloalkyl, which is optionally deuterated until fully deuterated;
  • n 0, 1, 2, 3, 4, 5 or 6;
  • R is a hydrophobic group
  • L 2 represents H or a solid support, or represents the position of connection with the adjacent nucleotide
  • R s , m and R are as defined in any one of technical solutions 1-5.
  • L 2 represents H or a solid support, or represents the position of connection with the adjacent nucleotide
  • H or a hydroxyl protecting group or represents the position of attachment to the adjacent nucleotide
  • the other represents H or a solid phase support, or indicates the position of connection with the adjacent nucleotide.
  • Technical Solution 15 An oligonucleotide according to any one of Technical Solutions 11-14, which comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at the 5’ end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • An oligonucleotide according to any one of Technical Solutions 11-15 which comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at its 3’ end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Technical Solution 17 An oligonucleotide according to any one of Technical Solutions 11-16, which comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at its 5’ end and 3’ end, respectively, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Technical Solution 18 An oligonucleotide according to any one of Technical Solutions 11-17, which contains one or more compounds of formula (I’) according to any one of Technical Solutions 11-13, or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, inside the oligonucleotide.
  • An oligonucleotide which comprises two or more hydrophobic groups inside the oligonucleotide, at the 5' end and/or the 3' end; preferably, the hydrophobic group is as defined as the R group in the compound of formula (I); preferably, the hydrophobic group is connected to the oligonucleotide via a linker, such as a biodegradable linker.
  • L 2 represents H or a hydroxyl protecting group, or represents the position for connection with the adjacent nucleotide
  • Rs is selected from H, D, halogen, C1-6 alkyl or C1-6 haloalkyl, which is optionally deuterated until fully deuterated;
  • n 0, 1, 2, 3, 4, 5 or 6;
  • R is a hydrophobic group
  • L 2 represents H or a hydroxyl protecting group, or represents the position for connection with the adjacent nucleotide
  • R s , m and R are as defined in any one of technical solutions 1-5.
  • L 2 represents H or a hydroxyl protecting group, or represents the position for connection with the adjacent nucleotide
  • R s , m and R are as defined in technical solutions 1-5.
  • One of them represents H, or represents the position of connection with the adjacent nucleotide, and the other indicates H or a hydroxyl protecting group, or the position of attachment to the adjacent nucleotide.
  • Technical Solution 23 A double-stranded RNA according to any one of Technical Solutions 20-22, wherein the positive strand comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at the 5’ end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Technical Solution 24 A double-stranded RNA according to any one of Technical Solutions 20-23, wherein the positive strand comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at the 3’ end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Technical Solution 25 A double-stranded RNA according to any one of Technical Solutions 20-24, wherein the positive strand comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at the 5’ end and the 3’ end, respectively, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Technical Solution 26 A double-stranded RNA according to any one of Technical Solutions 20-25, wherein the positive strand comprises one or more compounds of formula (I’) according to any one of Technical Solutions 11-13, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, inside the oligonucleotide.
  • Technical Solution 28 A double-stranded RNA according to any one of Technical Solutions 20-27, wherein the antisense strand comprises a compound of formula (I’) according to any one of Technical Solutions 11-13 at the 3’ end, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Technical Solution 30 A double-stranded RNA according to any one of Technical Solutions 20-29, wherein the antisense strand comprises one or more compounds of formula (I’) according to any one of Technical Solutions 11-13, or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, inside the oligonucleotide.
  • Technical solution 31 A double-stranded RNA according to any one of technical solutions 20-30, wherein two or more of the sense strand and/or antisense strand There are at least 5 to 30 nucleotides between each compound of formula (I'), or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof.
  • Double-stranded RNA according to any one of Technical Solutions 20-31, wherein the double-stranded RNA comprises two compounds of formula (I’), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, which are located at any two of the following sites: the 5’ end of the sense strand, the 3’ end of the sense strand, the 5’ end of the antisense strand and the 3’ end of the antisense strand; preferably located at the 5’ end of the sense strand and the 3’ end of the sense strand.
  • formula (I’) pharmaceutically acceptable salts, tautomers or stereoisomers thereof
  • a double-stranded RNA according to any one of Technical Solutions 20 to 32, wherein the double-stranded RNA comprises three compounds of formula (I’), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, which are located at any three of the following sites: the 5’ end of the sense strand, the 3’ end of the sense strand, the 5’ end of the antisense strand and the 3’ end of the antisense strand; preferably located at the 5’ end of the sense strand, the 3’ end of the sense strand and the 3’ end of the antisense strand.
  • formula (I’) pharmaceutically acceptable salts, tautomers or stereoisomers thereof, which are located at any three of the following sites: the 5’ end of the sense strand, the 3’ end of the sense strand, the 5’ end of the antisense strand and the 3’ end of the antisense strand; preferably located at the 5’ end of the sense strand, the 3’ end of the sense strand and the 3’ end
  • Double-stranded RNA according to any one of Technical Solutions 20-33, wherein the double-stranded RNA comprises four compounds of formula (I’), or pharmaceutically acceptable salts, tautomers or stereoisomers thereof, which are located at the following sites: the 5’ end of the sense chain, the 3’ end of the sense chain, the 5’ end of the antisense chain and the 3’ end of the antisense chain.
  • the double-stranded RNA of any one of Technical Solutions 20-33 further comprising a terminal phosphate protecting group or a prodrug protecting group coupled to the 5' end of the antisense strand, preferably a vinyl phosphate ester group or a prodrug protecting group represented by formula (X):
  • X1 is selected from OH or
  • R a is selected from H, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl or C 2-6 alkynyl, which is optionally deuterated until fully deuterated;
  • R b and R c are independently selected from H, C 1-6 alkyl or C 1-6 haloalkyl, and said R b and R c may be optionally substituted by D, C 6-10 aryl or 5-10 membered heteroaryl until fully deuterated;
  • X2 is a chemical bond connected to the first nucleotide at the 5' end of the antisense strand, preferably connected through a hydroxyl group;
  • X3 is independently selected from O or S;
  • T is selected from
  • each R T1 is independently selected from H, D, halogen, CN, C 1-6 alkyl , C 1-6 haloalkyl, C 1-6 haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, or a chain comprising GalNAc, which is optionally deuterated, up to fully deuterated;
  • Each R T2 is independently selected from H, D, halogen, CN, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl or C 2-6 alkynyl, which is optionally deuterated until fully deuterated;
  • Each R T3 is independently selected from H, D, halogen, CN, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl or C 2-6 alkynyl, which is optionally deuterated until fully deuterated;
  • Each R T4 is independently selected from H, D, halogen, CN, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl or C 2-6 alkynyl, which is optionally deuterated until fully deuterated;
  • n 0, 1, 2, 3, 4 or 5;
  • n 0, 1, 2, 3, 4 or 5;
  • p 0, 1, 2, 3, 4 or 5;
  • X is selected from a chemical bond, -O-, -S-, -C(O)-, -C(O)O-, -OC(O) -, -OC(O)NRX1-, -NRX1C(O)O-, -NRX1C ( O )- or -C(O) NRX1- ;
  • RX1 is selected from H, C1-6 alkyl or C1-6 haloalkyl, which is optionally deuterated until fully deuterated;
  • L is -Ar-(CH 2 ) 1-6 -O-, wherein each CH 2 may be optionally substituted by R#, R# is selected from H, D, halogen, CN, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl or C 2-6 alkynyl, which is optionally deuterated until fully deuterated;
  • Ar in L is connected to X, and the oxygen atom is connected to the phosphorus atom;
  • Ar is selected from C 3-10 cycloalkyl, 3-10 membered heterocyclyl, C 6-10 aryl or 5-14 membered heteroaryl, wherein the C 3-10 cycloalkyl, 3-10 membered heterocyclyl, C 6-10 aryl or 5-10 membered heteroaryl may be optionally substituted with 1, 2, 3, 4 or 5 R*;
  • R* is selected from H, D, halogen, CN, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl or C 2-6 alkynyl, which is optionally deuterated up to fully deuterated;
  • P1 is selected from a protecting group, preferably a hydroxy protecting group, such as trimethylsilyl (TMS), triethylsilyl (TES), dimethylisopropylsilyl (DMIPS), diethylisopropylsilyl (DEIPS), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS), acetyl (Ac), chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl (TFA), benzoyl, p-methoxybenzoyl, 9-fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Allo c), 2,2,2-trichloroethoxycarbonyl (Troc), benzyloxycarbonyl (Cbz), tert-butyloxy
  • the double-stranded RNA of any one of Technical Solutions 20-35 which is selected from small interfering RNA (siRNA) and short hairpin RNA (shRNA), and is preferably used to inhibit genes expressed in the eye.
  • siRNA small interfering RNA
  • shRNA short hairpin RNA
  • LL6, LL7 and LL8 are selected from the following compounds:
  • nucleic acid chain means connected to the 3' carbon or corresponding position of the previous nucleotide or nucleotide analog via a phosphate group, a phosphorothioate group or other linking group, It means that it is connected to the 5' carbon or corresponding position of the next nucleotide or nucleotide analog through a phosphate group, a thiophosphate group or other linking group; when the corresponding structure is located at the terminal position of the nucleic acid chain, Correspondingly, this refers to attachment to hydrogen, terminal modifications, terminal protecting groups, or other structures that may be used at the ends of nucleic acid chains.
  • Technical solution 40 A vector comprising a nucleotide sequence encoding the double-stranded RNA described in any one of the aforementioned technical solutions 20-39.
  • Technical solution 41 A cell containing the double-stranded RNA as described in any one of Technical Solutions 20-39 or the vector as described in Technical Solution 40.
  • Technical Solution 42 A pharmaceutical composition comprising the double-stranded RNA as described in any one of Technical Solutions 20-39, the vector as described in Technical Solution 40, or the cell as described in Technical Solution 41, and optionally a pharmaceutically acceptable carrier or excipient.
  • Technical solution 43 A kit comprising the double-stranded RNA as described in any one of Technical Solutions 20-39, the vector as described in Technical Solution 40, or the cell as described in Technical Solution 41.
  • nucleic acid chain means connected to the 3' carbon or corresponding position of the previous nucleotide or nucleotide analog via a phosphate group, a phosphorothioate group or other linking group, It means that it is connected to the 5' carbon or corresponding position of the next nucleotide or nucleotide analog through a phosphate group, a phosphorothioate group or other linking group; when the corresponding structure is located at the terminal position of the nucleic acid chain, Correspondingly, this refers to attachment to hydrogen, terminal modifications, terminal protecting groups, or other structures that may be used at the ends of nucleic acid chains.
  • oligonucleotides are as follows, wherein the sequence of 5'->3' from the to connect.
  • the corresponding structure is located in the middle of the nucleic acid chain, means connected to the 3' carbon or corresponding position of the previous nucleotide or nucleotide analog via a phosphate group, a phosphorothioate group or other linking group, It means that it is connected to the 5' carbon or corresponding position of the next nucleotide or nucleotide analog through a phosphate group, a thiophosphate group or other linking group; if the corresponding structure is located at the terminal position of the nucleic acid chain, Correspondingly, this refers to attachment to hydrogen, terminal modifications, terminal protecting groups, or other structures that may be used at the ends of nucleic acid chains.
  • the organic phase was washed with a 1:1 mixed solution of saturated NaHCO 3 and saturated saline (20 mL x 3), and then the organic phase was dried over Na 2 SO 4 and spun dry.
  • the crude product was purified by prep-HPLC (chromatographic column: Waters Xbridge BEH C18 150*25mm*5um; mobile phase: TEAA-ACN; gradient: 55%-95%/16min; flow rate: 15ml/min) to give a yellow oily compound DL0082 (170mg, yield 35.34%, purity 97.26%).
  • reaction solution was spin-dried, and the crude product was purified by prep-HPLC (column: 01-Waters Xbridge BEH C18 19 ⁇ 150 mm, 5 ⁇ m; conditions: TEAA-ACN; Begin B 65-95; gradient time: 15 minutes; 100% B holding time: 2 minutes; flow rate: 15 ml/min) to obtain a yellow oily product DL0084 (80.0 mg, yield: 36.27%).
  • the combined organic phase was washed with ammonium chloride solution (20.0 mL x 1), sodium bicarbonate solution (20.0 mL x 1), and saturated brine (20.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product.
  • reaction solution was dried by spin drying, and the crude product was purified by prep-HPLC (column: Waters Xbridge BEH C18 100*25mm*5um; condition: TEAA-ACN; Begin B 55-95; gradient time: 15 minutes; 100% B holding time: 6 minutes; flow rate: 15 ml/min), to obtain a yellow oily product DL0127 (120 mg, 0.133 mmol, 35.56%).
  • the reaction solution was diluted with DCM (50.0 mL), washed with saturated citric acid aqueous solution (10.0 mL x 3), the organic phase was washed with saturated NaHCO 3 aqueous solution (20.0 mL x 3) and saturated saline aqueous solution (20.0 mL x 3), and then the organic phase was dried with Na 2 SO 4 , and the organic phase was spin-dried to obtain a crude product.
  • the reaction solution was diluted with DCM (20.0 mL), washed with saturated citric acid aqueous solution (5.00 mL x 3), saturated NaHCO 3 aqueous solution (5.00 mL x 3) and saturated saline solution (5.00 mL x 3), and then the organic phase was dried over anhydrous Na 2 SO 4 , and the organic phase was spin-dried to obtain a crude product.
  • the reaction solution was diluted with dichloromethane (30 mL) and washed with saturated citric acid solution (15 mL x 3), saturated sodium bicarbonate solution (15 mL x 3) and saturated sodium chloride solution (15 mL x 3) in sequence.
  • the product 3 was obtained as a white solid (630 mg, yield 30.87%).
  • reaction solution was spin-dried, and the crude product was purified by prep-HPLC (column: Waters Xbridge BEH C18 100*25mm*5um; conditions: TEAA-CAN; Begin B 55-95; gradient time: 15 minutes; 100% B holding time: 6 minutes; flow rate: 15 ml/min) to obtain a yellow oily product DL0135 (140 mg, yield 31.91%, purity 99.54%).
  • the reaction solution was diluted with ethyl acetate (50 mL) and washed with saturated citric acid solution (30 mL x 3), saturated sodium bicarbonate solution (30 mL x 3) and saturated sodium chloride solution (30 mL x 3) in sequence.
  • the product 3 was obtained as a white solid (1530 mg, yield 81.02%).
  • reaction solution was diluted with dichloromethane (10 mL), and washed with saturated citric acid solution (10 mLX3), saturated sodium bicarbonate solution (10 mLX3) and saturated sodium chloride solution (10 mLX3) in sequence.
  • the crude product was purified by Prep-HPLC (chromatographic column: Waters Xbridge BEH C18 100*25mm*5um; mobile phase: TEAA-ACN; gradient: 65%-95%/15min; flow rate: 15ml/min).
  • a colorless oily product DL0136 (163mg, yield 40.63%, purity 97.57%) was obtained.
  • the crude product was purified by prep-HPLC (chromatographic column: Waters Xbridge BEH C18 150*25mm*5um; mobile phase: TEAA-ACN; gradient: 65%-95%/15min; flow rate: 15ml/min) to obtain a white solid compound DL0137 (45.0mg, yield 33.69%, purity 95.06%).
  • the crude product was prepared and separated by a reverse phase column (chromatographic column: Waters Xbridge BEH C18 100*25mm*5um; mobile phase: TEAA-CAN; B%: 60%-95%, 10min; flow rate: 15ml/min) to obtain a colorless oily liquid DL0138 (178 mg, yield 52.93%, purity 97.55%).
  • reaction solution was diluted with dichloromethane (50.0 mL), washed with citric acid (10 mL x 3), sodium bicarbonate (10 mL x 3), and saturated sodium chloride (10 mL x 3), and the organic phase was dried over anhydrous sodium sulfate, filtered and dried to obtain a yellow solid product compound 3 (250 mg, 0.613 mmol, 86.00%).
  • aqueous phase was extracted with dimethyltetrahydrofuran (20 mL x 3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and dried to obtain a brown solid product, compound 4 (350 mg, 0.889 mmol, 109.84%).
  • reaction solution was diluted with dichloromethane (100 mL), washed with saturated sodium bicarbonate aqueous solution (30 mL x 3) and saturated sodium chloride aqueous solution (30 mL x 3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a brown solid crude product compound 6 (600 mg, 0.727 mmol).
  • reaction solution was spin-dried, and the crude product was purified by prep-HPLC (column: Waters Xbridge BEH C18 100*25mm*5um; conditions: TEAA-ACN; Begin B 55-95; gradient time: 15 minutes; 100% B holding time: 6 minutes; flow rate: 15 ml/min) to obtain a yellow oily product DL0140 (120 mg, yield 35.89%, purity 97.34%).
  • reaction solution was dried by spin drying, 10.0 mL of water was added, the pH of the aqueous phase was adjusted to ⁇ 4 with 1M HCl, the aqueous phase was extracted with dimethyltetrahydrofuran (10.0 mL x 2), the combined organic phase was washed with saturated brine (10.0 mL x 1), dried over anhydrous sodium sulfate, filtered, and spin dried to obtain a white solid crude product 7 (460 mg, 0.55 mmol, 73.6%).
  • reaction solution was spin-dried, and the crude product was purified by prep-HPLC (column: Waters Xbridge BEH C18 19*150 mm; conditions: TEAA-ACN; Begin B 65-95; gradient time: 15 minutes; 100% B holding time: 2 minutes; flow rate: 15 ml/min) to obtain a white solid product DL0142 (106 mg, 0.08 mmol, 53.9%).
  • reaction liquid was directly suspended under reduced pressure to obtain a crude product, which was prepared and separated by a reverse phase column (chromatographic column: 01-Waters Xbridge BEH C18 19*150 mm; mobile phase: TEAA-ACN; gradient: 75%-95%/18 minutes; flow rate: 15 ml/minute) to obtain a colorless oily compound DL0143 (110 mg, yield 55.49%, purity 99.49%).
  • a reverse phase column chromatographic column: 01-Waters Xbridge BEH C18 19*150 mm; mobile phase: TEAA-ACN; gradient: 75%-95%/18 minutes; flow rate: 15 ml/minute
  • the reaction solution was diluted with DCM (50.0 mL), washed with saturated NaHCO 3 aqueous solution (20.0 mL x 3), the organic phase was washed with saturated saline solution (20.0 mL x 3), and then the organic phase was dried with Na 2 SO 4 , and the organic phase was spin-dried to obtain a crude product.
  • the reaction solution was diluted with DCM (50.0 mL), washed with saturated NaHCO 3 aqueous solution (20.0 mL X 3), and the organic phase was washed with saturated saline solution (20.0 mL X 3), and then the organic phase was dried with Na 2 SO 4 , and the organic phase was spin-dried to obtain a crude product.
  • the reaction solution was diluted with DCM (20.0 mL), washed with saturated citric acid aqueous solution (5.00 mL X 3), saturated NaHCO 3 aqueous solution (5.00 mL X 3) and saturated saline solution (5.00 mL X 3), and then the organic phase was dried over anhydrous Na 2 SO 4 , and the organic phase was spin-dried to obtain a crude product.
  • Compound 8 500 mg).
  • the crude product was prepared and separated by a reverse phase column (chromatographic column: 01-Waters Xbridge BEH C18 19*150 mm; mobile phase: TEAA-ACN; B%: 75%-95%, 15 min; flow rate: 15 ml/min) to obtain a white solid compound DL0144 (218 mg, yield 65.91%, purity 91.17%).
  • the crude product was purified by Prep-HPLC (chromatographic column: Waters Xbridge BEH C18 100*25mm*5um; mobile phase: TEAA-ACN; gradient: 45%-95%/16min; flow rate: 15ml/min) to obtain a colorless oily product DL0145 (84.0mg, yield 36.26%, purity 98.70%).
  • the siRNA of the present invention is prepared using the solid phase phosphoramidite method well known in the art.
  • the specific method can be referred to, for example, PCT Publication Nos. WO2016081444 and WO2019105419, and is briefly described as follows.
  • nucleoside monomers are connected one by one from the 3'-5' direction according to the arrangement order of the positive chain nucleotides.
  • Each connection of a nucleoside monomer includes four steps of deprotection, coupling, capping, oxidation or thiolation, and the synthesis scale is 5umol of oligonucleotides.
  • the synthesis conditions are as follows:
  • the nucleoside monomer was provided in a 0.05 mol/L acetonitrile solution, and the reaction conditions for each step were the same, i.e., the temperature was 25 degrees.
  • a 3% trichloroacetic acid-dichloromethane solution was used for deprotection, and the deprotection was repeated 3 times.
  • the activating agent used in the coupling reaction was a 0.25 mol/L 5-ethylthiotetrazolyl (ETT)-acetonitrile solution, and the coupling was repeated 2 times.
  • ETT 5-ethylthiotetrazolyl
  • the capping reaction used 10% acetic anhydride-acetonitrile and pyridine/N-methylimidazole/acetonitrile (10:14:76, v/v/v) and the capping was repeated 2 times.
  • the oxidation reaction used 0.05 mol/L iodine in tetrahydrofuran/pyridine/water (70/20/10, v/v/v) and the oxidation was repeated 2 times.
  • the thiolation reaction used 0.2 mol/L phenylacetyl disulfide (PADS) in acetonitrile/3-methylpyridine (1/1, v/v) and the thiolation was repeated 2 times.
  • PADS phenylacetyl disulfide
  • nucleoside monomers are connected one by one from the 3'-5' direction according to the arrangement order of the antisense chain nucleotides.
  • Each connection of a nucleoside monomer includes four steps of deprotection, coupling, capping, oxidation or thiolation.
  • the synthesis conditions of 5umol oligonucleotides of the antisense chain are the same as those of the sense chain.
  • a column filled with strong anion fillers can be used, and a sodium chloride-sodium hydroxide system can be used for elution and purification, and the product can be collected and piped.
  • a gel filler purification column can be used for desalination, and the elution system is pure water.
  • Example 3 In vivo activity testing (CNS delivery)
  • A, U, G and C represent the natural adenine ribonucleotide, uracil ribonucleotide, guanine ribonucleotide and cytosine ribonucleotide, respectively.
  • d indicates that the adjacent nucleotide on the right is a deoxyribonucleotide.
  • dA, dT, dG, and dC represent adenine deoxyribonucleotide, thymine deoxyribonucleotide, guanine deoxyribonucleotide, and cytosine deoxyribonucleotide, respectively.
  • i inosine ribonucleotide
  • m indicates that the adjacent nucleotide on its left is a 2'-OCH 3 modified nucleotide.
  • Am, Um, Gm and Cm represent 2'-OCH 3 modified A, U, G and C.
  • f indicates that the adjacent nucleotide on its left side is a 2'-F modified nucleotide.
  • Af, Uf, Gf, and Cf represent 2'-F modified A, U, G, and C, respectively.
  • s indicates that the two adjacent nucleotides and/or delivery vectors are linked by phosphorothioate.
  • VP indicates that the adjacent nucleotide on the right is a vinyl phosphate-modified nucleotide.
  • Ib represents an inverted abasic deoxyribonucleotide, which may include the following three structures depending on its position/connection mode in siRNA.
  • mice SD rats, male, 6-8 weeks old, 2-3 rats per group;
  • the residual inhibition rate was calculated by the following formula:
  • ⁇ Ct [(target gene in Ct experimental group-internal reference in Ct experimental group)-(target gene in Ct control group-internal reference in Ct control group)].
  • the target gene was SOD1
  • the internal reference was GAPDH
  • the control group was injected with artificial cerebrospinal fluid (aCSF).
  • DR005713, DR005714, DR005715, DR005717, DR005718, DR005716, and DR005735 all reduced the expression of SOD1 in the central nervous system.
  • the knockdown effect of DR005735 was better than that of other sequences.
  • Example 4 In vivo activity testing (ocular delivery)
  • sequence information used in this example is as follows:
  • C57BL/6 mice male, 6-8 weeks were randomly divided into groups and administered a single dose of 2 ⁇ g per eye by bilateral intravitreal injection.
  • the siRNA conjugate was administered in a 5 mg/mL solution (phosphate buffer solution as solvent); specifically, before the experiment, the siRNA conjugate was dissolved in phosphate buffer solution and diluted to the required solution concentration and volume, and the administration volume of phosphate buffer solution and siRNA conjugate was 1.5 ⁇ L/eye.
  • the eyeballs were removed and separated into three parts: 1 retinal pigment epithelium (RPE) + choroid + sclera; 2 retina; 3 cornea + iris + ciliary body; the separated samples were immediately frozen in liquid nitrogen and then stored at -80°C for detection of mTTR mRNA.
  • RPE retinal pigment epithelium
  • the 2 - ⁇ Ct value was calculated and converted into a percentage to obtain the residual inhibition rate
  • ⁇ Ct [(target gene in Ct experimental group-internal reference in Ct experimental group)-(target gene in Ct control group-internal reference in Ct control group)].
  • the target gene was mTTR and the internal reference was mGAPDH.

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Abstract

L'invention concerne un composé de formule (I) ou un sel, un tautomère ou un stéréoisomère pharmaceutiquement acceptable de celui-ci. Le composé de formule (I) est contenu à l'intérieur d'un nucléotide, ou à une extrémité 5' et/ou à une extrémité 3', et est utilisé pour améliorer la capacité d'un ARN double brin à passer à travers la barrière hémato-encéphalique. La présente invention concerne en outre un kit de test, une composition pharmaceutique, une cellule, un support, un ARN double brin ou un oligonucléotide comprenant le composé de formule (I).
PCT/CN2023/127728 2022-10-31 2023-10-30 Ligand pour administrer de l'arnsi à l'oeil et au système nerveux central WO2024093907A1 (fr)

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Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4252801A (en) * 1980-01-04 1981-02-24 E. R. Squibb & Sons, Inc. Morpholinyl acetamide derivatives and use thereof
WO2004080406A2 (fr) * 2003-03-07 2004-09-23 Alnylam Pharmaceuticals Compositions therapeutiques
WO2004094595A2 (fr) * 2003-04-17 2004-11-04 Alnylam Pharmaceuticals Inc. Agents modifiés d'arni
CN101679324A (zh) * 2007-06-15 2010-03-24 田边三菱制药株式会社 吗啉衍生物
CN104640847A (zh) * 2012-09-14 2015-05-20 上海医药集团股份有限公司 新型肾素抑制剂
WO2015095276A1 (fr) * 2013-12-19 2015-06-25 Merck Sharp & Dohme Corp. Inhibiteurs de la protéase du vih
WO2015134366A1 (fr) * 2014-03-06 2015-09-11 Merck Sharp & Dohme Corp. Inhibiteurs de la protéase du vih
WO2022084331A2 (fr) * 2020-10-20 2022-04-28 Sanofi Nouveaux ligands pour le récepteur d'asialoglycoprotéine
WO2023143571A1 (fr) * 2022-01-30 2023-08-03 大睿生物医药科技(上海)有限公司 Ligand de ciblage contenant de la n-acétylgalactosamine

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4252801A (en) * 1980-01-04 1981-02-24 E. R. Squibb & Sons, Inc. Morpholinyl acetamide derivatives and use thereof
WO2004080406A2 (fr) * 2003-03-07 2004-09-23 Alnylam Pharmaceuticals Compositions therapeutiques
WO2004094595A2 (fr) * 2003-04-17 2004-11-04 Alnylam Pharmaceuticals Inc. Agents modifiés d'arni
CN101679324A (zh) * 2007-06-15 2010-03-24 田边三菱制药株式会社 吗啉衍生物
CN104640847A (zh) * 2012-09-14 2015-05-20 上海医药集团股份有限公司 新型肾素抑制剂
WO2015095276A1 (fr) * 2013-12-19 2015-06-25 Merck Sharp & Dohme Corp. Inhibiteurs de la protéase du vih
WO2015134366A1 (fr) * 2014-03-06 2015-09-11 Merck Sharp & Dohme Corp. Inhibiteurs de la protéase du vih
WO2022084331A2 (fr) * 2020-10-20 2022-04-28 Sanofi Nouveaux ligands pour le récepteur d'asialoglycoprotéine
WO2023143571A1 (fr) * 2022-01-30 2023-08-03 大睿生物医药科技(上海)有限公司 Ligand de ciblage contenant de la n-acétylgalactosamine

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE REGISTRY 29 April 2021 (2021-04-29), ANONYMOUS: "- 4-Morpholinecarboxylic acid, 2,6-bis(hydroxymethyl)-, 1,1-dimethylethyl ester, (2S,6S)- (CA INDEX NAME)", XP093166465, Database accession no. 2639392-38-2 *
DATABASE REGISTRY 6 December 2015 (2015-12-06), ANONYMOUS: "- 4-Morpholinecarboxylic acid, 2-(hydroxymethyl)-6-[(phenylmethoxy)methyl]-, 1,1-dimethylethyl ester (CA INDEX NAME)", XP093166467, Database accession no. 1823598-00-0 *
QIAN CHEN: "(2 R *,6 S *)- tert -Butyl 2,6-bis(hydroxymethyl)morpholine-4-carboxylate", ACTA CRYSTALLOGRAPHICA SECTION E, JOHN WILEY & SONS, GB, vol. 66, no. 7, 15 July 2010 (2010-07-15), GB , pages o1623 - o1623, XP093166441, ISSN: 1600-5368, DOI: 10.1107/S1600536810017599 *

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